低能量激光对成熟脂肪细胞去分化的实验研究

Experimental study on the dedifferentiation of mature adipocytes with low-level laser

目的探讨低能量激光对成熟脂肪细胞发生去分化及前脂肪细胞数量的影响。方法应用天花板培养法培养脂肪细胞,贴壁后将培养的脂肪细胞随机分为对照组与照射组。照射组应用低能量激光进行照射,照射时间5min,连续3d;对照组不给予激光照射。应用倒置显微镜观察细胞形态变化,采用免疫荧光染色法检测2组脂肪前体细胞标志物CD29表达情况。结果对照组细胞贴壁后第14天出现细胞脱脂滴甚至不含脂滴现象,而照射组细胞贴壁后第6天即出现细胞脱脂滴甚至不含脂滴现象;对照组球形细胞多,胞质内充满大量折光性脂滴,显示出近似成熟脂肪细胞的形态;照射组细胞逐渐变小,梭形与不规则细胞明显增多,其中圆形空泡状细胞去分化为成纤维细胞状,胞质内见少量细小脂滴或不含脂滴,有明显向前脂肪细胞去分化的表现;对照组细胞CD29表达为阴性,照射组细胞CD29表达为阳性。结论低能量激光照射后人成熟脂肪细胞会发生去分化改变,变成类似前脂肪细胞的细胞。

Objective To investigate the influence of low-level laser on dedifferentiation of mature adipocytes and the preadipocyte number. Methods Human subcutaneous mature adipocyte was cultured by using lattice culturing method. After adhesion, the adipocytes were randomly divided into radiation group and control group. Radiation group received low-level laser radiation for 5 minutes, totally for 3 days, and control group received no radiation. The cellular morphologic changes were observed 24, 48 and 72 hours after radiation respectively under inverted microscope. The expression of CD29 was detected by immunofluorescence staining. Results After mature adipocytes firmly adhering to the wall, the lipid droplets of control group in the cytoplasm were gradually released and extruded out the cells from the 14th day, even no lipid droplets were found in the cytoplasm. The same phenomenon of radiation group occurred from the 6th day. The spherocytes increased in number, with a large number of reflective lipid droplets inside the cytoplasm in the form of mature adipocytes in control group. In radiation group, the cells became small, spindle-shaped and irregular cells increased in number obviously, in which the round vacuolated cells were dedifferentiated into fibroblast-like cells with few tiny lipid droplets inside the cytoplasm showing obvious dedifferentiation into preadipocytes. The expression of CD29 was negative in control group and positive in radiation group. Conclusion Low-level laser could dedifferentiate the human mature adipocyte into preadipocyte-like cell.